Putative target relationship between miR-26a and toll-lke receptor (TLR)3 in rats. (A) Schematic diagram of pairing relationship between miR-26a and tlr3 mRNA 3′UTR in rats. The rat tlr3 mRNA 3′UTR sequence contains a putative binding site of miR-26a analyzed by the bioinformatics software . (B) Effect of miR-26a mimics/inhibitor on the luciferase activity of pMIR-TLR3 vector. Target relationship between tlr3 mRNA 3′UTR and miR-26a was analyzed by dual luciferase reporter assay, pMIR-REPORT™ Luciferase vectors with or without tlr3 mRNA 3′UTR element (pMIR-TLR3 or pMIR empty vectors) were transfected into Hela cells, and pRL-TK vector was used as an internal control reporter in all conditions for normalization. NC, negative control; pMIR, pMIR-REPORT™ Luciferase vector. Bars represent standard error of the mean (SEM) from three transfections. Experiments were independently repeated twice. *Statistically significant difference (P <0.05) compared with NC miRNA; ^significant difference compared with vector (Mann–Whitney U-test). (C) Effect of miR-26a mimics on the luciferase activity of wild-type or mutated pMIR-TLR3 vector. pMIR-REPORT™ Luciferase vector carrying a TLR3 mRNA 3′UTR element with the putative binding site of miR-26a was transfected into Hela cells (upper panel). Bars represent SEM from three experiments. *Statistically significant difference (P <0.05, Mann–Whitney U-test). Schematic diagram of pairing relationship between miR-26a and wild-type or mutated pMIR-TLR3 vector indicates that three nucleotides have been altered in the mutated pMIR-TLR3 vector (lower panel).